Antimicrobial endolysin polypeptides, compositions and formulations

An anti-microbial, endolysin technology, applied in the field of host cells, can solve the problems of affecting pathogenic bacteria, adverse effects, etc.

Pending Publication Date: 2021-11-19
阿希坦有限公司
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, antibiotic treatment not only affects pathogenic bacteria but also adversely affects beneficial and protective flora in the chicken gut

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Antimicrobial endolysin polypeptides, compositions and formulations
  • Antimicrobial endolysin polypeptides, compositions and formulations
  • Antimicrobial endolysin polypeptides, compositions and formulations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 13

[0286] 1. Inoculate Clostridium perfringens strain NCTC 8237 into BHI+C medium, and culture at 37°C under anaerobic conditions overnight to the stationary phase;

[0287] 2. Inoculate the overnight culture into fresh BHI+C medium, and culture it at 37°C under anaerobic conditions until the OD600nm is about 0.6;

[0288] 3. Centrifuge 1.8 mL of the culture at 16600×g for 5 minutes at room temperature under anaerobic conditions;

[0289] 4. Remove the supernatant, and resuspend the cell pellet in 0.9 mL resuspension buffer (NaCl127mM, NaCl1 2 HPO 4 70mM, NaH 2 PO 4 30mM pH 7.0);

[0290] 5. Dilute the cell suspension to OD 600nm 0.6 with resuspension buffer under anaerobic conditions;

[0291] 6. Add the purified endolysin polypeptide or fragment to a concentration of 5 μg / mL under anaerobic conditions;

[0292] 7. Incubate at 25°C for 1 hour under anaerobic conditions;

[0293] 8. Determine viable cell counts by serial dilution, plating cells on RCM solid medium, and i...

Embodiment 12

[0307] 1. Inoculate Clostridium perfringens strain NCTC 8237 into BHI+C medium, and culture at 37°C under anaerobic conditions overnight to the stationary phase;

[0308] 2. Inoculate the overnight culture into fresh BHI+C medium and culture under anaerobic conditions at 37°C until the culture reaches exponential phase (OD 620nm about 1.0);

[0309] 3. Centrifuge a certain volume of exponential phase culture at 3800×g for 30 minutes at 10°C, then remove the supernatant;

[0310] 4. Resuspend the cells in a volume of PBS (pH 7.0) that is half of the initial pre-centrifuged culture volume to concentrate the cells;

[0311] 5. Transfer 90 μL of resuspended cells to a well of a 96-microwell plate, and add 10 μL of a 10-fold concentrated endolysin polypeptide or fragment in PBS (pH 7.0) to a final concentration of 5 μg / mL;

[0312] 6. Monitor the decrease in turbidity by taking OD 620nm kinetic readings at appropriate intervals on a microplate reader at room temperature;

[0313]...

Embodiment 15

[0320] 1. Inoculate Clostridium perfringens strain Cp6 into BHI+C medium, and culture at 37°C under anaerobic conditions overnight to the stationary phase;

[0321] 2. Dilute the stationary phase culture in fresh LB medium under anaerobic conditions;

[0322] 3. Under anaerobic conditions, prepare serial dilutions of the purified endolysin stock solution in PBS (pH 7.0) to a concentration 10 times higher than the final desired protein loading concentration;

[0323] 4. Under anaerobic conditions, mix serial dilutions of purified endolysin with diluted stationary phase cultures in wells of a microtiter plate, where, for each well, mix 45 μL of diluted stationary phase cultures with 5 μL of 10-fold concentrated and purified endolysin was mixed to a final cell load of approximately 10 4 cells / mL;

[0324] 5. Incubate the cells overnight at 41°C for 12-20 hours under anaerobic conditions;

[0325] 6. Determine the degree of growth inhibition by measuring the change in OD on a m...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention relates to novel endolysin polypeptides or fragments thereof which possess antimicrobial activity, preferably antibacterial activity against Clostridium perfringens. The invention also relates to a nucleic acid molecule encoding the endolysin polypeptide or fragment, a recombinant polynucleotide expression vector comprising such a nucleic acid molecule, as well as a host cell comprising such a nucleic acid molecule or comprising such a recombinant polynucleotide expression vector. The invention also relates to compositions and foodstuffs comprising the endolysin polypeptides or fragments and uses thereof in the treatment of diseases or disorders in animals.

Description

technical field [0001] The present invention relates to novel endolysin polypeptides or fragments thereof having antimicrobial activity, preferably antibacterial activity against Clostridium perfringens. The present invention also relates to a nucleic acid molecule encoding the endolysin polypeptide or fragment, a recombinant polynucleotide expression vector comprising the nucleic acid molecule, and a host cell comprising the nucleic acid molecule or the recombinant polynucleotide expression vector. The present invention also relates to compositions and foodstuffs comprising the endolysin polypeptides or fragments and their use in the treatment of diseases or disorders in animals. Background technique [0002] Concerns about antibiotic resistance have led to public and regulatory pressure to ensure their judicious use in human and veterinary medicine. In food animal production, the prevention of animal diseases especially caused by Clostridium perfringens has become a bigge...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/36C12N9/80A23L3/3571A23B4/22C11D3/48
CPCC12N9/2462C12N9/80C11D3/48A23K20/189C11D3/38636A23K10/12A23K50/75A23K50/30A23K50/50C12N2330/50
Inventor 李维洛K·米勒F·J·纳瓦罗
Owner 阿希坦有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap